This SBIR Phase II project will develop a cost-effective, easy-to-use, field-portable, and sensitive instrument for rapid screening of avian influenza viruses (AIVs). The Phase I research has demonstrated the feasibility of rapid and sensitive detection of H5N1 AIV in a buffer solution by combination of patent pending bionanobead sample pretreatment, capillary immunocapture, and nano-biolabeling technologies. The proposed biosensor technology was shown to be 103- 104 times more sensitive than a commercial rapid test strip. In phase II, the research will be directed to develop a commercial prototype for a cross-contamination-free biosensor system for rapid detection of AIVs and/or H5/H7 subtypes in swab samples. The target detection limit is a 4-log increase in sensitivity as compared to commercially available rapid test strips. This level of sensitivity will allow for determination of the presence of AIV in asymptomatic birds and will therefore be a useful screen for AI at an early stage of infection. The H5N1 highly pathogenic AIV is the world's largest current pandemic threat. H5 and H7 strains can mutate from low pathogenicity to high pathogenicity during replication and circulation in poultry and thus are very dangerous. Consequently, strengthening the surveillance of AI infection and development of rapid screening technology for in-field detection of AIVs and H5/H7 subtypes is urgently needed. Nearly 2 million AI tests are run each year on turkey and broiler production flocks. The proposed testing system has several advantages over AI testing that is performed in centralized laboratories. The advantages include reduced instrumentation price and cost per test, more rapid turn around time for test results, and reduced need for personnel with specialized training. The proposed system also has significant advantages over currently-approved rapid test kits because of its ability to detect both H5 and H7 subtypes and capability of effectively screening AI infection in seemingly healthy birds prior to shipment for harvest. PUBLIC HEALTH RELEVANCE: The proposed biosensor technology will greatly improve the ability to identify the presence of influenza A and the notifiable subtypes (H5 and H7) of AI in birds that are showing signs of illness as well as in birds that are asymptomatic. This product will be valuable to the poultry industry as it will allow for implementation of an early warning system to minimize the spread of the disease and prevent poultry products contaminated with AIV from entering the food supply. The regulatory and research community will also be able to monitor the presence of the disease in wild birds in order to gain a better understanding of the spread of this virus on local and global scales.